【摘要】 将不可渗透型的保护剂海藻糖有效地载入血小板内部是用冷冻干燥法保存血小板重要的第一步。研究血小板对海藻糖的载入量随外部海藻糖浓度、孵化时间、孵化温度改变的变化规律,发现在细胞外海藻糖浓度为50mmol/L、孵化温度37℃、孵化时间4h的条件下,血小板能有效地吸收海藻糖,细胞内海藻糖浓度达到15mmol/L以上。对孵化后的血小板进行形态观察、血液学分析和膜联蛋白(annexin)V结合活化分析,结果表明孵化后的血小板保持了正常血小板的形态和功能。更多还原

【Abstract】 Introducing the impermeable lyoprotectant trehalose into the cytoplasm is the first step toward preserving platelets by freeze-drying. We investigated the loading of the human platelets with trehalose. Trehalose uptake was measured as a function of extracellular trehalose concentration, incubation time and incubation temperature. We found an effective uptake of the trehalose at 37 ℃ in a time span of 4 h in the presence of 50 mmol/L external trehalose. The internal trehalose concentration reached above 15 mmol/L. Morphological study, hematology analysis and annexin V binding assay showed that trehalose loaded platelets maintain the normal morphology and function of platelets. These data provide the foundation in developing a protocol for freeze-drying platelets.更多还原